Overview

The LANCE® Ultra Europium-anti-di/mono-methyl-Histone H3 Lysine 27 (H3K27me2-1) antibody was used for the development and optimization of a EZH2 methyltransferase assay using a biotinylated Histone H3 (21-44) as substrate. A JMJD3 demethylase assay was also developed, using a biotinylated Histone H3 (21-44) peptide tri-methylated at lysine 27 as substrate. A technical note describing the assay is available in our product literature.

Epigenetic enzymatic assays are optimized using a biotinylated histone H3-derived peptide as substrate. The modified peptide is captured by the Eu-labeled antibody (Ab) and ULight-Streptavidin (SA) which bring the Eu donor and ULight acceptor dye molecules into close proximity. Upon irradiation at 320 or 340 nm, the energy from the Eu donor is transferred to the ULight acceptor dye which, in turn, generates light at 665 nm. The intensity of the light emission is proportional to the level of biotinylated substrate modification.